Evidence map›Paper›PMID 41787538›Full record

ArticleBMC research notes2026

Standardization of C6/36 cells for DENV-2 virological assays: isolation, TCID₅₀ and serum neutralization.

Elmo José Dos Santos, Amanda da Rocha Santos, Mayanna Moreira Costa Fogaça, Naiara Soares da Silva, Milena Silva Souza, Ruth Dálety da Silva Brito, Paloma Oliveira Vidal, Samuel Santos Pereira, Jéssica Pires Farias, Alexander Birbrair and 4 more

Abstract read
PubMed Full text at PMC Publisher
In one paragraph

Article in BMC research notes, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 1 paper.

0numbers the graph read from it
0cells of the map it votes in
1citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

1 citing paper in PubMed.

  1. Article
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

14 authors.

Elmo José Dos SantosWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Amanda da Rocha SantosWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Mayanna Moreira Costa FogaçaWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Naiara Soares da SilvaWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Milena Silva SouzaWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Ruth Dálety da Silva BritoWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Paloma Oliveira VidalWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil.
Samuel Santos PereiraVaccine Development Laboratory, Department of Microbiology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil.
Jéssica Pires FariasVaccine Development Laboratory, Department of Microbiology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil.
Alexander BirbrairDepartment of Dermatology, School of Medicine and Public Health, University of Wisconsin-Madison, Madison, WI, USA.
Rúbens Prince Dos Santos AlvesInstitut Pasteur de São Paulo, São Paulo, Brazil.
Wilson Barros LuizDepartment of Biological Sciences, State University of Santa Cruz, Ilhéus, Bahia, Brazil.
Luís Carlos de Souza FerreiraVaccine Development Laboratory, Department of Microbiology, Biomedical Sciences Institute, University of São Paulo, São Paulo, Brazil.
Jaime Henrique AmorimWestern Bahia Virology Institute, Center of Biological Sciences and Health, Federal University of Western Bahia, Barreiras, Bahia, Brazil. jaime.amorim@ufob.edu.br.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

objectiveThis study aimed the standardization of detection, isolation, titration, replication kinetics, and virus neutralization assay protocols for DENV-2 (New Guinea C strain) with the C6/36 mosquito-derived cell line.

resultsDENV-2 was isolated, titrated by TCID₅₀, and quantified via molecular assays using serially diluted viral RNA to generate a standard curve using the C6/36 cell line. The serum neutralization assay was optimized in 96-well plates based on visual scoring of cytopathic effect (CPE). Optimized serum dilutions effectively reduced nonspecific cytotoxicity caused by residual complement, preventing misinterpretation and enabling reliable detection of serum neutralizing activity in C6/36 cells. RT-qPCR assays calibrated using a TCID₅₀-titrated DENV-2 viral stock showed high amplification efficiency (90–100%) with R² ≥ 0.98. The results demonstrate that the standardized workflow enables proof-of-principle DENV-2 neutralization testing in C6/36 cells, yielding neutralization patterns consistent with those observed in Vero cells, and supports the use of this platform as an alternative for resource-limited laboratories.

Indexed as

Antibodies, NeutralizingAntibodies, ViralDengue VirusNeutralization TestsAnimalsCell LineChlorocebus aethiopsRNA, ViralVero CellsVirus ReplicationAntibodies, NeutralizingAntibodies, ViralRNA, ViralDengue virusInsect cellsNeutralization titersStandardizationVirus titration

Identifiers

PMID41787538
PMCPMC13077881

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LicenceCC BY-NC-ND
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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.